Fetal hemoglobin induction by histone deacetylase inhibitors involves generation of reactive oxygen species

Cheng Hui Hsiao, Wei Li, Tzu Fang Lou, B. Surendra Baliga, Betty Sue Pace

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Objective. Several compounds, including butyrate and trichostatin A, have been shown to activate γ-gene expression via p38 mitogen-activated protein kinase (MAPK) signaling. In eukaryotic cells, reactive oxygen species (ROS) act as signaling molecules to mediate phosphorylation of tyrosine kinases such as p38 MAPK to regulate gene expression. Therefore, we determined the role of the reactive oxygen species hydrogen peroxide (H2O2) in drug-mediated fetal hemoglobin (HbF) induction. Methods. H2O 2 levels were measured using 2′,7′-dichlorofluorescein- diacetate in K562 cells after drug treatments. To confirm a role for H 2O2 in HbF induction, studies were completed with the mitochondrial respiratory chain inhibitor myxothiazole, which prevents ROS generation. The ability of myxothiazole to block γ-globin mRNA accumulation and HbF induction was measured in K562 cells and burst-forming unit-erythroid colonies respectively using quantitative real-time PCR and alkaline denaturation. Results. Butyrate and trichostastin A stimulated p38 MAPK phosphorylation via a H2O2-dependent mechanism. Pretreatment with myxothiazole to inhibit ROS formation or SB203580 to impede p38 MAPK signaling attenuated γ-gene activation in K562 cells and HbF induction in erythroid progenitors. However, myxothiazole had no effect on the ability of hydroxyurea to induce HbF. Conclusion. The findings presented herein support a ROS-p38 MAPK cell signaling mechanism for HbF induction by butyrate and trichostatin A.

Original languageEnglish (US)
Pages (from-to)264-273
Number of pages10
JournalExperimental Hematology
Volume34
Issue number3
DOIs
StatePublished - Mar 1 2006
Externally publishedYes

Fingerprint

Fetal Hemoglobin
Histone Deacetylase Inhibitors
p38 Mitogen-Activated Protein Kinases
Reactive Oxygen Species
K562 Cells
Butyrates
trichostatin A
Phosphorylation
Gene Expression
Erythroid Precursor Cells
Hydroxyurea
Globins
Eukaryotic Cells
Electron Transport
Pharmaceutical Preparations
Protein-Tyrosine Kinases
Hydrogen Peroxide
Transcriptional Activation
Real-Time Polymerase Chain Reaction
Messenger RNA

ASJC Scopus subject areas

  • Molecular Biology
  • Hematology
  • Genetics
  • Cell Biology
  • Cancer Research

Cite this

Fetal hemoglobin induction by histone deacetylase inhibitors involves generation of reactive oxygen species. / Hsiao, Cheng Hui; Li, Wei; Lou, Tzu Fang; Baliga, B. Surendra; Pace, Betty Sue.

In: Experimental Hematology, Vol. 34, No. 3, 01.03.2006, p. 264-273.

Research output: Contribution to journalArticle

Hsiao, Cheng Hui ; Li, Wei ; Lou, Tzu Fang ; Baliga, B. Surendra ; Pace, Betty Sue. / Fetal hemoglobin induction by histone deacetylase inhibitors involves generation of reactive oxygen species. In: Experimental Hematology. 2006 ; Vol. 34, No. 3. pp. 264-273.
@article{b2d0315497814c85935be2bfdf008c72,
title = "Fetal hemoglobin induction by histone deacetylase inhibitors involves generation of reactive oxygen species",
abstract = "Objective. Several compounds, including butyrate and trichostatin A, have been shown to activate γ-gene expression via p38 mitogen-activated protein kinase (MAPK) signaling. In eukaryotic cells, reactive oxygen species (ROS) act as signaling molecules to mediate phosphorylation of tyrosine kinases such as p38 MAPK to regulate gene expression. Therefore, we determined the role of the reactive oxygen species hydrogen peroxide (H2O2) in drug-mediated fetal hemoglobin (HbF) induction. Methods. H2O 2 levels were measured using 2′,7′-dichlorofluorescein- diacetate in K562 cells after drug treatments. To confirm a role for H 2O2 in HbF induction, studies were completed with the mitochondrial respiratory chain inhibitor myxothiazole, which prevents ROS generation. The ability of myxothiazole to block γ-globin mRNA accumulation and HbF induction was measured in K562 cells and burst-forming unit-erythroid colonies respectively using quantitative real-time PCR and alkaline denaturation. Results. Butyrate and trichostastin A stimulated p38 MAPK phosphorylation via a H2O2-dependent mechanism. Pretreatment with myxothiazole to inhibit ROS formation or SB203580 to impede p38 MAPK signaling attenuated γ-gene activation in K562 cells and HbF induction in erythroid progenitors. However, myxothiazole had no effect on the ability of hydroxyurea to induce HbF. Conclusion. The findings presented herein support a ROS-p38 MAPK cell signaling mechanism for HbF induction by butyrate and trichostatin A.",
author = "Hsiao, {Cheng Hui} and Wei Li and Lou, {Tzu Fang} and Baliga, {B. Surendra} and Pace, {Betty Sue}",
year = "2006",
month = "3",
day = "1",
doi = "10.1016/j.exphem.2005.12.009",
language = "English (US)",
volume = "34",
pages = "264--273",
journal = "Experimental Hematology",
issn = "0301-472X",
publisher = "Elsevier Inc.",
number = "3",

}

TY - JOUR

T1 - Fetal hemoglobin induction by histone deacetylase inhibitors involves generation of reactive oxygen species

AU - Hsiao, Cheng Hui

AU - Li, Wei

AU - Lou, Tzu Fang

AU - Baliga, B. Surendra

AU - Pace, Betty Sue

PY - 2006/3/1

Y1 - 2006/3/1

N2 - Objective. Several compounds, including butyrate and trichostatin A, have been shown to activate γ-gene expression via p38 mitogen-activated protein kinase (MAPK) signaling. In eukaryotic cells, reactive oxygen species (ROS) act as signaling molecules to mediate phosphorylation of tyrosine kinases such as p38 MAPK to regulate gene expression. Therefore, we determined the role of the reactive oxygen species hydrogen peroxide (H2O2) in drug-mediated fetal hemoglobin (HbF) induction. Methods. H2O 2 levels were measured using 2′,7′-dichlorofluorescein- diacetate in K562 cells after drug treatments. To confirm a role for H 2O2 in HbF induction, studies were completed with the mitochondrial respiratory chain inhibitor myxothiazole, which prevents ROS generation. The ability of myxothiazole to block γ-globin mRNA accumulation and HbF induction was measured in K562 cells and burst-forming unit-erythroid colonies respectively using quantitative real-time PCR and alkaline denaturation. Results. Butyrate and trichostastin A stimulated p38 MAPK phosphorylation via a H2O2-dependent mechanism. Pretreatment with myxothiazole to inhibit ROS formation or SB203580 to impede p38 MAPK signaling attenuated γ-gene activation in K562 cells and HbF induction in erythroid progenitors. However, myxothiazole had no effect on the ability of hydroxyurea to induce HbF. Conclusion. The findings presented herein support a ROS-p38 MAPK cell signaling mechanism for HbF induction by butyrate and trichostatin A.

AB - Objective. Several compounds, including butyrate and trichostatin A, have been shown to activate γ-gene expression via p38 mitogen-activated protein kinase (MAPK) signaling. In eukaryotic cells, reactive oxygen species (ROS) act as signaling molecules to mediate phosphorylation of tyrosine kinases such as p38 MAPK to regulate gene expression. Therefore, we determined the role of the reactive oxygen species hydrogen peroxide (H2O2) in drug-mediated fetal hemoglobin (HbF) induction. Methods. H2O 2 levels were measured using 2′,7′-dichlorofluorescein- diacetate in K562 cells after drug treatments. To confirm a role for H 2O2 in HbF induction, studies were completed with the mitochondrial respiratory chain inhibitor myxothiazole, which prevents ROS generation. The ability of myxothiazole to block γ-globin mRNA accumulation and HbF induction was measured in K562 cells and burst-forming unit-erythroid colonies respectively using quantitative real-time PCR and alkaline denaturation. Results. Butyrate and trichostastin A stimulated p38 MAPK phosphorylation via a H2O2-dependent mechanism. Pretreatment with myxothiazole to inhibit ROS formation or SB203580 to impede p38 MAPK signaling attenuated γ-gene activation in K562 cells and HbF induction in erythroid progenitors. However, myxothiazole had no effect on the ability of hydroxyurea to induce HbF. Conclusion. The findings presented herein support a ROS-p38 MAPK cell signaling mechanism for HbF induction by butyrate and trichostatin A.

UR - http://www.scopus.com/inward/record.url?scp=33644881627&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33644881627&partnerID=8YFLogxK

U2 - 10.1016/j.exphem.2005.12.009

DO - 10.1016/j.exphem.2005.12.009

M3 - Article

VL - 34

SP - 264

EP - 273

JO - Experimental Hematology

JF - Experimental Hematology

SN - 0301-472X

IS - 3

ER -